Seasonal variations in the sporulating capacity of the fungus causing coffee berry disease

Assessments of the sporulating capacity of Colletotrichum coffeanum Noack in coffee branches (defined as the number of conidia produced per cm²per hour under optimum conditions) have been made for 4 years on a number of different sites in Kenya. An annual rhythm is evident which is essentially similar on all sites and for all years. Early-season and late-season peaks are separated by a mid-season trough. The change in sporulating capacity at any time is negatively correlated with wetness in the immediately preceding period, and daily rainfall can be used as a measure of wetness. The position of the peaks and their duration is defined by the timing of the onset and the duration of the monsoon rains, and is, therefore, subject to considerable variation. It is suggested that the correct timing of control sprays might be related to the form of the annual curve of sporulating capacity, and be predictable from rainfall records.     

The stimulating effect of some fungicides on Glomerella cingulata in relation to the control of coffee berry disease

The non-parasitic strain of Colletotrichum coffeanum Noack which colonizes the maturing wood of Coffea arabica produces its perfect stage (Glomerella cingulata) on the branches. Assessments of the relative abundance of this in relation to preceding rainfall have shown that for unsprayed branches, but rarely for sprayed ones, the correlation is negative and highly significant. Various fungicides stimulate G. cingulata greatly, and this stimulation persists for many months. One other saprophytic component of the micro–flora is similarly stimulated, but with two others all the fungicides used depress sporulation. It is suggested that when the main source of infection of coffee berries is the pathogenic conidia from the wood, the mechanism of long–term control by early–season sprays may lie partly in this stimulation of some fungal competitors of the pathogen.     

Rainfall and cropping patterns in relation to coffee berry disease*

Both the increase in severity and extent of coffee berry disease (CBD) during the 1960s and the failure of early-season sprays to control it have been attributed to changes in climatic conditions and cropping patterns. A critical analysis of rainfall data since 1951, when CBD first appeared in the East Rift coffee districts of Kenya, produces no evidence that conditions affecting CBD development were any different in the 1960s than in the 1950s, in either high-or low-altitude coffee-growing areas. A marked increase in overlapping crops occurred during the late 1950s, and this was caused mainly by a change in pruning methods. The increased disease hazard from this change of cropping pattern cannot account for the failure of early season sprays, which were successful before 1962. An estimate of the time of main crop flowering shows that only in those years when flowering was early and the rainy season finished early did early-season sprays achieve substantial benefits. It is concluded that their effect in these years was obtained by fungicidal protection of the crop throughout most of the rainy season, and that their failure in other years contributed to the worsening disease situation.     

Evaluation of fungicides for the control of coffee berry disease in Kenya

Some sixty fungicides were tested in the laboratory for ability to ( a ) depress sporulation of Colletotrichum coffeanum on the maturing bark of coffee shoots and ( b ) inhibit spore germination. On the basis of these tests over twenty products were selected for testing in the field during 1968-70. Nine of these (50 % copper formulations, Ortho-Difolatan, Daconil, Benlate, Topsin, Top-sin M, Tecto 60, Delan and Du-Ter) could be recommended for control of coffee berry disease (CBD) in Kenya.
Field evaluation of fungicides is complicated by multiple flowering which creates a succession of overlapping crops, and by high variability in cropping potential which is often aggravated by normal pruning practices; in some circumstances other diseases and pests may also be a complicating factor. As a result, yield, which should be the most sensitive measure of CBD control, is at times even less satisfactory than disease estimates based on a small sample of the developing crop. Disease incidence in the ripe crop, which is easy to measure, is unfortunately not always well correlated with disease and crop loss earlier in the season. Consequently, critical assessment of fungicide performance requires estimates of both disease progress and crop loss.
Consideration of fungicide performance indicates that physical characteristics, particularly persistence and capacity for redistribution, are especially important for effective control of CBD.     

A comparison of fungicide spray volumes for the control of coffee berry disease

A continuation of previous trials in which three sprayers were used to apply 8.4 g. of Perenox in a range of spray volumes from 0.2 to 1.7 l. per tree confirmed that the volume of spray did not affect greatly the deposit retained on the branch surface. Branches were able to retain up to about 250 mg. Cu/m.² and it is considered that a volume equivalent to 100 gal./acre would provide the greatest margin of safety to compensate for inaccuracies of application, while avoiding serious run-off. Where coffee was well pruned, intensive copper spraying raised the annual yield by about 2 cwt. of clean coffee per acre. Copper spraying did not provide an adequate control of coffee berry disease and, under certain circumstances, increased incidence.     

Some factors influencing the performance of spray programmes for the control of coffee berry disease

In 1968 in a field trial at Kamundu, Kenya, early-season (Jan.-Mar.) sprays of copper, captafol and Tuzet failed to control coffee berry disease (CBD), in the late (main) crop of cultivars S.L.28 and S.L.34. Copper and captafol caused some delay in disease development but eventually infection reached over 50 % in all treatments and final yields of sprayed plots were below those from unsprayed plots. In a similar trial in 1969 at Yara on the cultivar French Mission a similar spray programme achieved significant benefits: copper and captafol kept the level of disease below 20% and gave improved yields compared with unsprayed plots. A continuation of the sprays until June increased yields still further. In a second trial in 1968 at Kiamara on the cultivar French Mission substantial quantities of early crop were present and disease in this was effectively controlled by early-season sprays of captafol, although the late crop on the same trees became severely affected. Removal of this overlapping early crop in January caused only a limited delay in disease development in the late crop, despite the fact that with some treatments this stripping resulted in the removal of a large source of potential inoculum from the tree. The results are considered in relation to differences between seasons and fungicides and it is concluded that further evidence has been provided for the importance of the protective role of fungicides against CBD. It is considered that stripping of the early crop is never likely to be a necessary management practice.     

In vitro screening of coffee genotypes for resistance to coffee berry disease (Colletotrichum kahawae)

An in vitro protocol was developed to screen Coffea arabica genotypes for resistance to coffee berry disease caused by Colletotrichum kahawae. Initially, cultural conditions which influenced the growth of isolates of C. kahawae on agar media suitable for callus growth were determined. The growth of the fungus on the callus derived from susceptible and resistant genotypes was then assessed. This ensured that no detrimental competition for nutrients between the pathogen and the calli occurred. Optimisation of the concentration of the phytohormones added to the media, the temperature and incubation period were found to be important in the expression of differential responses of calli to inoculation with the pathogen as detected by measurement of hyphal growth. The screening of calli of nine C. arabica genotypes showed that this method identified genotypes highly resistant or susceptible to the disease and was sufficiently sensitive to distinguish those genotypes with moderate or low resistance.     

The development of the grape berry cuticle in relation to susceptibility to bunch rot disease

Some physical and morphological factors of grape berry cuticlewere investigated at different developmental stages of threeclones ofVitis vinifera cv. Pinot noir. The surface morphologyof grape berries was examined by scanning electron microscopyand cuticle anatomy was examined by light and transmission electronmicroscopy. During the period from flowering to maturity, thecomposition of the cuticular waxes changed, corresponding withan increase of waxy deposits and significant modifications ofthe wax surface morphology. The content in cutin per unit surfacedecreased more than 2.5-fold between berry set (16 d after anthesis)and veraison of the grape berries, and might predispose thegrape berry to fungal infection. This result was correlatedwith the differentiation of the cuticle layers and particularlywith a decrease in the thickness of the primary cuticle at harvest. Key words: Botrytis cinerea, cuticle, cutin, epicuticular waxes, Vitis vinifera L     

Strains of Colletotrichum coffeanum, the causal agent of coffee berry disease, tolerant to benzimidazole compounds in Kenya

The detection of Colletotrichum coffeanum tolerant to methyl ester of benzimidazole 2-carbamic acid (carbendazim) and a related benzimidazole compound, cypendazole, followed increases in levels of coffee berry disease observed on Coffea arabica in experimental plots sprayed for 2 yr with these compounds. Sporulation by the pathogen on naturally infected berries removed from carbendazim-, cypendazole- or benomyl-sprayed plots was not checked by a further application of 0–05 % (a.i.) of any of the compounds. Nearly all the isolates from these berries were capable of some growth on agar media containing 1000 ppm (a.i.) of either carbendazim or cypendazole. However, only a few could tolerate 1000 ppm of benomyl and the inability of this compound to reduce sporulation on berries infected with tolerant strains was presumably due to its rapid conversion to carbendazim within the host tissue. Less than 1 ppm of carbendazim, cypendazole or benomyl was needed to give 50% inhibition of conidia of the normal strain. Against the most tolerant strains, however, the LD 50 was > 100 ppm of carbendazim and about 30 ppm of benomyl. Whether isolated from unsprayed or benzimidazole-sprayed plots, all isolates of Colletotrichum acutatum, a saprophytic cohabitant of lesions initiated on berries by C. coffeanum, showed the highest degree of tolerance to benzimidazole compounds. No tolerance of either fungus to the ‘conventional’ fungicide captafol was detected.     

Current status of coffee berry disease (Colletotrichum kahawae Waller & Bridge) in Ethiopia

The current status of coffee berry disease (CBD) caused by Colletotrichum kahawae was intensively assessed and examined in 152 sample coffee farms from 22 districts across major coffee growing regions of Ethiopia. The results showed that CBD was prevalent with significantly (p < 0.001) varied intensity of damage among fields, districts and zones. The highest disease incidence of 70.7, 65.3 and 59.3% was recorded in Hararghe, Gedeo and Jimma, with correspondingly higher severity of 42.7, 46.7 and 32.0%, respectively. The national average incidence and severity of CBD was 52.5 and 29.9% that indicated the present status of the disease is remarkably on increasing trend. The increased intensity of CBD was strongly associated with reduced disease management practices (r = 0.50), altitude (r = 0.42), coffee cultivars (r = 0.23) and production systems (r = 0.28). This empirical evidence shows that CBD is on an upsurge and remains a major challenge to Arabica coffee production in Ethiopia.     

Effect of bilirubin on the fungicidal capacity of human neutrophils

The effect of unconjugated bilirubin on the fungicidal capacity of human neutrophils was examined. Per cent Torulopsis glabrata killed in the presence of 1 x 10(-5) and 5 x 10(-5) M bilirubin was 72.7 +/- 2.5 and 40.9 +/- 7.2 respectively, compared to 81.4 +/- 4.1 in control (p less than 0.01 and p less than 0.001), respectively). This effect was not due to a concomitant inhibition of phagocytosis. The results suggest that jaundiced neonates may be more susceptible to fungal infections.     

The effect of acid shock on sporulating Bacillus subtilis cells

AIMS: To study the effect of acid shock in sporulation on the production of acid-shock proteins, and on the heat resistance and germination characteristics of the spores formed subsequently. METHODS AND RESULTS: Bacillus subtilis wild-type (SASP-alpha+beta+) and mutant (SASP-alpha-beta-) cells in 2 x SG medium at 30 degrees C were acid-shocked with HCl (pH 4, 4.3, 5 and 6 against a control pH of 6.2) for 30 min, 1 h into sporulation. The D85-value of B. subtilis wild-type (but not mutant) spores formed from sporulating cells acid-shocked at pH 5 increased from 46.5 min to 78.8 min, and there was also an increase in the resistance of wild-type acid-shocked spores at both 90 degrees C and 95 degrees C. ALA- or AGFK-initiated germination of pH 5-shocked spores was the same as that of non-acid-shocked spores. Two-dimensional gel electrophoresis showed only one novel acid-shock protein, identified as a vegetative catalase 1 (KatA), which appeared 30 min after acid shock but was lost later in sporulation. CONCLUSIONS: Acid shock at pH 5 increased the heat resistance of spores subsequently formed in B. subtilis wild type. The catalase, KatA, was induced by acid shock early in sporulation, but since it was degraded later in sporulation, it appears to act to increase heat resistance by altering spore structure. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first proteomic study of acid shock in sporulating B. subtilis cells. The increasing spore heat resistance produced by acid shock may have significance for the heat resistance of spores formed in the food industry.     

Epicatechin and catechin may prevent coffee berry disease by inhibition of appressorial melanization of Colletotrichum kahawae

Colletotrichum kahawae is the causal agent of coffee berry disease. Appressorial melanization is essential for the fungal penetration of plant cuticle. Epicatechin is abundant in green coffee berry pericarp. Inoculation of C. kahawae conidial suspension containing 1.2 mg epicatechin or catechin/ml did not affect conidial germination or appressorial formation but appressorial melanization was completely inhibited and infection by the treated conidia was less than 30% of the untreated control. Epicatechin and catechin may, therefore, prevent coffee berry disease by inhibition of the appressorial melanization of C. kahawae.     

Altitude and coffee production systems influence extent of infestation and bean damage by the coffee berry borer

The coffee berry borer (CBB), Hypothenemus hampei Ferrari (Coleoptera: Curculionidae: Scolytinae), is one of the major insect pests of coffee worldwide. The present study was designed to assess the level of infestation of coffee berries at different developmental stages across different altitudes and coffee management systems. The experiment was carried out at three locations in southwestern Ethiopia under two coffee management systems and four coffee berry development stages with three replications. Results of the study showed significantly highest proportion of damaged berries (37.5%), number of holes per berry (10.88) and number of adult CBB per berry (7.55) on dried leftover berries at low-altitude study sites. On the other hand, the lowest mean percent damaged berries, number of holes per berry and number of adults were recorded at mid- and high-altitude study sites. The study also showed that, CBB caused significantly highest damage in plantation coffee management system than garden coffee. Results of this study highlight proper harvesting at red ripe stage in order to minimise incidence of CBB. It is also important to design integrated management strategies to mitigate CBB damage especially in lowland plantation coffee production systems.     

Diversity of Bacillus thuringiensis strains isolated from coffee plantations infested with the coffee berry borer Hypothenemus hampei

The coffee berry borer Hypothenemus hampei Ferrari (Coleoptera: Scolytidae) was first reported infecting Costa Rican coffee plantations in the year 2000. Due to the impact that this plague has in the economy of the country, we were interested in seeking new alternatives for the biological control of H. hampei, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 202 B. thuringiensis isolates obtained from Costa Rican coffee plantations infested with H. hampei were analyzed through crystal morphology of the crystal inclusions and SDS-PAGE of 6-endotoxins, while 105 strains were further evaluated by PCR for the presence cry, cyt and vip genes. Most of the Bt strains showed diverse crystal morphologies: pleomorphic (35%), oval (37%), bipyramidal (3%), bipyramidal and oval (12%), bipyramidal, oval and pleomorphic (10%) and bipyramidal, oval and cubic (3%). The SDS-PAGE analyses of the crystal preparations showed five strains with delta-endotoxin from 20 to 40 kDa, six from 40 to 50 kDa, seven from 50 to 60 kDa, 19 from 60 to 70 kDa, 29 from 70 to 100 kDa and 39 from 100-145 kDa. PCR analyses demonstrated that the collection showed diverse cry genes profiles having several genes per strain: 78 strains contained the vip3 gene, 82 the cry2 gene, 45 the cry1 and 29 strains harbored cry3-cry7 genes. A total of 13 strains did not amplified with any of the cry primers used: cry1, cry2, cry3-7, cry5, cry11, cry12 and cry14. Forty-three different genetic profiles were found, mainly due to the combination of cry1A genes with other cry and vip genes. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against H. hampei and other insect pests of agricultural importance.     

The influence of weather on the seasonal incidence of coffee berry moth,Prophantis smaragdina (Butler)

The seasonal incidence of the coffee berry moth, Prophantis smaragdina (Butler) (Lepidoptera: Pyralidae), was investigated on Coffea arabica L. in Jimma, Ethiopia. Our results showed that; the coffee berry moth was present throughout the study period except during November and December. The average incidence was 24.5% and the peak incidence (61%) was in September. Multiple correlation analysis was used to estimate the strength of association between weather variables and the incidence of the pest, and stepwise (both forward and backward) regression analysis was used to select the best explanatory variable. There were strong associations among the explanatory weather variables, indicating the potential problem of multicollinearity in the regression analysis. Relative humidity had a highly significant regression coefficient of 2.228 and was selected in the stepwise regression analysis as the best explanatory variable. The results of the study can be used in designing an integrated pest management strategy against the coffee berry moth.     

A review of the biology and control of the coffee berry borer, Hypothenemus hampei (Coleoptera: Scolytidae)

The coffee berry borer, Hypothenemus hampei Ferrari, is a serious problem for the majority of the world's coffee growers and has proved to be one of the most intractable of present day pests. Despite a great deal of research, control still depends largely on the application of the organochlorine insecticide endosulfan, which is damaging to the environment, or a series of cultural and biological control methods which give variable and unpredictable results. This review summarizes the most important aspects of the biology and ecology of H. hampei and its control and identifies weak points in the knowledge about this pest. Emphasis is placed upon an analysis of the non-chemical control methods available and suggestions are offered for novel ecological and environmental factors worthy of further research, in the search for viable and sustainable control methods.     

Micro-meteorological studies of coffee berry disease A field investigation into the incidence of the physical conditions favourable for spore germination

In an investigation in a coffee plantation, the frequency and duration of the micro-meteorological conditions necessary for the germination of conidia of Colletotrichum coffeanum were evaluated on coffee twigs of different ages, aspects and exposures. No consistent relationships between aspect and disease incidence were established, but there were indications that east-facing branches might be slightly more disease-prone than west-facing ones because of more rapid warming of the surface water films by morning insolation. The macro-climate of the plantation was measured by conventional meteorological instruments. Daytime air temperatures, measured inside a Stevenson screen between the rows of trees, were, on average, about 1 C° and 2·5 C° lower than those recorded on twig surfaces under wet and dry conditions, respectively.